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  • Nextera DNA Flex Library Prep - TWB

    Does anyone know the composition of the Tagment Wash Buffer in the Nextera DNA Flex Library Prep (20018704) protocol? Does it contain alcohol? PEG?

    TIA

  • #2
    I would check out the SMART-SEQ2 protocol. They did homebrew something similar. I vaguely remember SDS to denature the enzyme.

    Comment


    • #3
      Hi BexMultiplex,

      According to the Nextera Flex patent, TWB is:

      -100 mM Tris HCl pH 8.5
      -100 mM NaCL
      -0.1% Tween 20

      The present disclosure relates to methods, compositions, and kits for treating target nucleic acids, including methods and compositions for fragmenting and tagging nucleic acid (e.g., DNA) using transposome complexes bound to a solid support.


      We have a Tecan, and are going to be reducing RXN volumes down to 1/4, and at that point TWB becomes a limiting reagent. I will be trying my own eventually, but I have not used it yet personally myself, so i can't promise it will work

      If you are ever trying to find what a reagent is, looking at the patent (if you can find it) is probably the best way to get this kind of information. It also really helpful for understanding how a kit work when the protocol is vague.

      Hope this helps!

      Comment


      • #4
        Thanks @UCan'tBcereus, that's really helpful!

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